Effect of intranasal administration of norethisterone and progesterone on pituitary and gonadal function in adult male and female bonnet monkeys (Macaca radiata).

The effect of intranasal spray of norethisterone (NET) and progesterone (P) on serum testosterone (T) and total sperm count in adult male bonnet monkeys and the effect of NET spray on the menstrual cycle in female monkeys has been studied. Whereas NET spray resulted in a drastic decrease in serum T levels and sperm count, P spray caused a decrease in total sperm count only. Intranasal spray of NET in adult cycling female monkeys between days 5 and 14 resulted in shortening of the menstrual cycle; this could be ascribed to a decrease in serum follicle-stimulating hormone, luteinizing hormone, 17 beta-estradiol, and P levels. Nasal spray of only solvent in male or female monkeys, however, had no effect on any of the parameters tested.

Steroid metabolism by monkey and human spermatozoa.

Freshly ejaculated spermatozoa from monkey and human were washed and incubated with tritium labelled androgens or estradiol to study the pattern of spermatozoa steroid metabolism. When equal concentrations of steroid substrates were used for incubation, monkey and human spermatozoa showed very similar pattern of steroid conversion. Spermatozoa from both species converted testosterone mainly to androstenedione, but reverse conversion of androstenedione to testosterone was negligible. Estradiol-17 beta was converted mainly to estrone. The close similarity between the spermatozoa of monkey and men in their steroid metabolic pattern indicates that the rhesus monkey could be an useful animal model to study the effect of drugs on the metabolic pattern of human spermatozoa.

Pharmacokinetics of norethisterone following its oral administration and by spraying it nasally or sublingually to male bonnet monkeys.

Pharmacokinetic parameters of norethisterone (NET) were studied in eight adult male bonnet monkeys following the administration of a single dose of 300 ug. The animals were crossed over between the following three routes of administration: oral ingestion, nasal and sublingual spraying. The results indicate that NET was readily absorbed by all three routes but the Cmax and AUC of NET were significantly greater by the sublingual route. No significant difference in the t 1/2 alpha or t 1/2 beta was observed between the three routes. These findings suggest that the sublingual route offers the possibility of reducing the effective dose of NET, which is widely used for contraceptive purposes.

Reproductive endocrine effects of intranasal administration of norethisterone (NET) to women.

Four consecutive menstrual cycles were studied in six healthy parous women. A solvent mixture comprising propylene glycol:ethanol:water (3:3:4) was sprayed intranasally daily using a glass atomizer between days 5 and 24 of the first (control) menstrual cycle. NET was dissolved in the solvent and similarly administered at a daily dose of 100 mcg during the second and third menstrual cycles. Nasal sprays were not administered during the fourth post-treatment cycle. Blood samples were taken during four consecutive cycles between days 8 and 15 and again between days 20 and 24 of the cycle to estimate levels of estradiol (E2), FSH, LH and progesterone (P). These studies revealed that nasal sprays of NET were well accepted and that no adverse clinical effects or menstrual disturbances occurred. NET inhibited ovulation in one cycle. The E2-induced mid-cycle rise in FSH and LH was either suppressed or inhibited in nine out of the 12 treated cycles. P levels in three treated cycles were indicative of luteal inadequacy. These endocrine effects of NET persisted into the post-treatment cycle in two cases.

Scanning electron microscopy in the assessment of sperm morphology.

The advantages of using scanning electron microscope (SEM) vis-a-vis light microscope (LM) to assess sperm morphology was studied. The semen samples obtained from 15 fertile (group I) and 25 infertile (group II) men were processed by routine procedures for LM and SEM. The usually described sperm abnormalities were identifiable with greater resolution. Sperm abnormalities were significantly more in group II, as compared with group I (P less than 0.001) by both methods. Abnormalities of different regions, like thinning of midpiece, were scored higher in SEM than in LM. The abnormality of aggregation or retraction of mitochondrial sheath was clearly visible under SEM and missed by LM. Apart from detection of certain subtle abnormalities, SEM has advantages of rapid screening, accurate quantitation directly from the screen, zooming to higher magnification and automation.

Gamete intrafallopian transfer: A preliminary experience.

BACKGROUND: After in vitro fertilizationand embryo transfer for tubal infertility, gamete intrafallopian transfer has been introduced for patients with non-tubal infertility. However, the gametes need to be transferred in 2 to 5 minutes and the distance between the operating theatre and tissue culture laboratory delayed its introduction at our hospital. METHODS AND PATIENTS: To overcome this problem we designed a box in which gametes could be stored. Using gametes taken from this box and employing the standard technique, we achieved 5 pregnancies in 39 infertile women. RESULTS: From 41 treatment cycles, 39 women underwent oocyte retrieval. Five pregnancies were achieved of which 4 delivered live births at full term and 1 ended in abortion. Our first gamete intrafallopian transfer baby was born on 6 January 1988. CONCLUSION: The gamete intrafallopian transfer technique can be successfullyadapted for India.

Cellular and humoral pathways in the neuroendocrine regulation of reproductive function.

PIP: Studies carried out in recent years, including those conducted by the author on the rhesus monkey, which have added more information on the possible cellular and humoral pathways by which the neural and endocrine functions concerned with reproductive phenomena are integrated are reviewed. The possible involvement of the circumventricular structures, especially the tanycyte ependyma and the pineal gland in the detection of the endogenous levels of gonadal hormones, and the cerebrospinal fluid (CSF) in the transmission of hormonal information to the various parts of the brain indicated further areas of research to be explored. Such exploration should be conducted in diverse, yet complimentary, disciplines so that the whole spectrum of events known to occur in the neuroendocrine regulation of reproduction can be studied.^ieng

Serum levels of testosterone, cortisol, prolactin and bioactive luteinizing hormone in adult male rhesus monkeys following cage-restraint or anaesthetizing with ketamine hydrochloride.

The present studies were carried out to characterize and compare the acute effects of cage-restraint or administration of a mild anaesthetic on serum levels of testosterone (T) cortisol (C), prolactin (Prl) and bioactive luteinizing hormone (LH) in adult male rhesus monkeys. Serum T levels declined progressively while C levels increased in the 3 sequential blood samples obtained at 20 min intervals from restrained monkeys. Prl and LH levels in the serum remained unaltered. Following single or multiple injections of ketamine hydrochloride (10 mg/kg) serum T levels did not decline progressively as seen in the blood samples obtained from the cage-restrained, conscious monkeys. Serum C and Prl levels showed a progressive rise following anaesthetization. LH levels were not affected by the anaesthetic. The findings reported here are of pertinent relevance to the evaluation of acute effects of any experimental procedure aimed at altering circulating levels of the hormones studied.

Association of tritium-labelled estradiol, progesterone and 17 alpha-hydroxyprogesterone with the proteins of the serum and cerebrospinal fluid in the rhesus monkey.

The electrophoretic profiles of the proteins in the serum and cerebrospinal fluid (CSF) of the rhesus monkey were studied by polyacrylamide gel electrophoresis. The association of tritium labelled estradiol (3H-E), progesterone (3H-P) and 17 alpha-hydroxyprogesterone (3H-17OHP) with these proteins was studied under in vitro and in vivo conditions. The CSF was found to contain, besides albumin, at least 4 globulins with similar electrophoretic mobilities as those found in the serum. The electrophoretic mobilities of these globulins were Ralb 0.4, 0.6, 0.8 and 0.9. The electrophoretic profiles of the monkey serum and CSF proteins were found to be similar to those described for human serum and CSF. 3H-E and 3H-P were respectively associated with serum globulins of Ralb 0.4 and 0.8 both in the in vitro and in vivo studies. The respective Ralb of these 2 globulins were similar to the testosterone-estradiol binding globulin and cortisol binding globulin in human serum. 3H-17OHP was found to be associated with several globulins under both in vitro and in vivo conditions. In marked contrast to the in vitro studies, radioactive products were found to be associated with albumin in vivo. It is presumed that these products may be steroidal conjugates derived from the systemic metabolism of the administered steroids. No radioactivity was associated with the CSF protein after in vitro incubation with any of the three 3H-steroids. In the in vivo studies, however, radioactivity was associated with such of the CSF proteins that were similar to those found in the serum and with which the administered steroids were associated.

Ultrastructure of the clear cell lining the epididymal lumen of the rat.

2 types of clear cells can be distinguished in the rat epididymis. The Type I clear cell occurs in the middle segment and it is distinguished by a centrally located nucleus and the presence of lipid droplets. The Type II clear cell occurs in the terminal segment; it has a basally situated nucleus and does not have lipid droplets. Ultrastructural studies indicate that the clear cells carry out a dual function of absorbing substances from the epididymal lumen by pinocytosis and secreting products into it by merocriny. both these functions are not carried out simultaneously by a particular clear cell but at different times. The Type I clear cell appears to secrete a glycolipoprotein while the Type II clear cell seems to secrete glycoproteins.

Ultrastructure of the cortex in the human egg.

Two distinct types of cortical granules were discerned in the human oocyte. The first type, G1 granules, had a mean diameter of 350 nm, contained a uniformly compacted, electron-dense substance; these granules were probably synthesized even before the oocyte was aspirated from mature follicles and their contents were never found to be secreted in any of the preovulatory (immature), unfertilized and fertilized eggs examined. The G2 granules measured 450 nm (mean diameter), had a granular content and were found to be synthesized and secreted at all the stages of egg development studied. Endocytosis was also evident in the unfertilized and fertilized eggs. The secretion of the G1 cortical granules even before fertilization suggests that they may have an additional role, rather than merely contributing to the blockade of polyspermy.

Structure of rete testis, vas efferens, epididymis and vas deferens of langur monkey (Presbytis entellus entellus Dufresne).

Morphological and histological features of rete testis, vas efferens, epididymis and vas deferens were studied in the langur monkey. Tubular extensions of rete were located towards lateral side of the testis. Its epithelium comprises mostly of cuboidal cells with hyaline cytoplasm. Three to nine bundles of vas efferens, emerging below the cranial pole of the testis, were observed. Vas efferens epithelium comprises of ciliated and nonciliated cells. Epididymis could be divided into six zones on the basis of cytological features. Principal cells, basal cells, apical cells and intraepithelial lymphocytes were observed in the epididymal epithelium, but their number, shape, size and location of nuclei varied in different zones. Vas deferens epithelium comprises of principal cells, basal cells, apical cells and few intraepithelial lymphocytes. Epithelium is surrounded by lamina propria, longitudinal, circular and longitudinal muscle layers.

Day and night levels of hormones in male rhesus monkeys kept under controlled or constant environmental light.

The marked differences between day and night levels of testosterone, cortisol, prolactin and bioactive luteinizing hormone observed in adult male rhesus monkeys maintained at 12 h light: 12 h dark schedule are not changed by exposing monkeys to constant illumination for a period of up to 15 days. These findings suggest that the photoperiodicity of environmental light may not be the principal determinant for the occurrence of the diurnal rhythms observed in the circulating levels of the 4 hormones studied.

Pharmacokinetics of progesterone after its administration to ovariectomized rhesus monkeys by injection, infusion, or nasal spraying.

The pharmacokinetics of progesterone (dose: 10 microgram per animal) were studied in blood and cerebrospinal fluid of adult ovariectomized rhesus monkeys after the administration of the steroid as an intravenous injection, intravenous infusion (duration of infusion: 10 min), or nasal spray. The bioavailability of progesterone, in terms of area under the time--concentration curve and the maximal concentration in the two body fluids, was significantly higher when the steroid was infused or sprayed intranasally than when it was injected intravenously. The clearance of the steroid from the serum, as estimated by its elimination rate constant, elimination half-life, and total body clearance, did not differ for the three methods of administration. These findings suggest that the bioavailability of progesterone is enhanced by extending the duration over which the steroid is delivered into the hemic circulation.

Bioavailability of progesterone enhanced by intranasal spraying.

The bioavailability of progesterone (P) in terms of area under time-concentration curve and maximal concentration in the serum and cerebrospinal fluid was studied in adult ovariectomized rhesus monkeys following the administration of P as a nasal spray, i.v. or i.m. injections, nasal or eye drops. The bioavailability of P in both the body fluids was found to be considerably higher following its being sprayed intranasally.

Reproductive exocrine and endocrine profiles and their seasonality in male langur monkeys (Presbytis entellus entellus).

The reproductive exocrine and endocrine profiles in male langurs are reported with an emphasis on seasonality. The animals showed positive response to electroejaculation throughout the year. The sperm concentration varied between 10-383 x 10(6)/ejaculation with wide fluctuations all through the year. No appreciable changes in the motility and percent live sperm were observed throughout the year. The levels of seminal fructose and magnesium remained unchanged throughout the year, while acid phosphatase showed wide fluctuations. Citric acid showed elevation during February and March and LDH showed elevated levels during April and May. The annual range of serum testosterone was 6-34 nMol/l with a peak during July. Cortisol ranged between 575-1587 nMol/l and prolactin ranged between 107-900 mU/l. Wide fluctuations were observed in hormonal levels. No seasonality was exhibited in the seminiferous tubule diameter, nuclear diameter of Sertoli cells and Leydig cells, and the cholesterol, glycogen, and sialic acid contents of testis. None of the parameters studied have shown any correlation with season. The results suggest that the male langurs lack seasonality in their reproductive exocrine and endocrine profiles and thus could be used as model for research in human reproduction.